Calibration using the same sample being measured enables quantitative in-the-field diagnostics for complex biological samples.

Abstract Simultaneous detection of multiple analytes from a single sample (multiplexing), particularly when done at the point need, can guide complex decision-making without increasing required volume or cost per test. Despite recent advances, multiplexed analyte sensing still typically faces critical limitation measuring only one type molecule (e.g., small molecules nucleic acids) assay platform. Here, we address this bottleneck with customizable platform that integrates cell-free...

RNA regulators are powerful components of the synthetic biology toolbox. Here, we expand repertoire gene networks built from these by constructing a transcriptional negative autoregulation (NAR) network out small RNAs (sRNAs). NAR motifs core natural genetic networks, and known for reducing response time steady state signal. Here use cell-free transcription–translation (TX-TL) reactions computational model to design prototype sRNA constructs. Using parameter sensitivity analysis, simple set...

In the host innate immune system, various pattern recognition receptors (PRRs) recognize conserved pathogen-associated molecular patterns (PAMPs) and represent an efficient first line of defense against invading pathogens. Toll-like (TLRs) are a major class PRRs, which able to wide range PAMPs play central role in initiating responses. TLR21 is one non-mammalian TLRs identified some bird fish species.In present study, we reported cloning identification cDNA from head kidney common carp...

Electroporation is a basic yet powerful method for delivering small molecules (RNA, DNA, drugs) across cell membranes by application of an electrical field. It used many diverse applications, from genetically engineering cells to drug- and DNA-based vaccine delivery. Despite this broad utility, the high cost electroporators can keep approach out reach budget-conscious laboratories. To address need, we develop simple, inexpensive, handheld electroporator inspired derived common household...

The field of metabolic engineering has yielded remarkable accomplishments in using cells to produce valuable molecules, and cell-free expression (CFE) systems have the potential push even further. However, CFE still face some outstanding challenges, including endogenous activity that is poorly understood yet a significant impact on productivity. Here, we use metabolomics characterize temporal changes their constituent components, central carbon amino acid metabolism. We find while changing...

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have proven instrumental in genetically manipulating many eukaryotic and prokaryotic organisms. Despite its usefulness, a CRISPR system had yet to be developed for use Neisseria gonorrhoeae (Gc), bacterium that is the main etiological agent of gonorrhea infection. Here, we programmable IPTG-inducible Type I-C interference (CRISPRi) derived from commensal species lactamica as gene repression Gc. As opposed...

We develop mismatched fluorescent probes to monitor the long noncoding RNA (lncRNA) in living cells.

Field-deployable diagnostics based on cell-free systems have advanced greatly, but on-site quantification of target analytes remains a challenge. Here we demonstrate that Escherichia coli lysate-based biosensors coupled to personal glucose monitor (PGM) can enable analyte quantification, with the potential for straightforward reconfigurability diverse types analytes. We show analyte-responsive regulators transcription and translation modulate production reporter enzyme β-galactosidase, which...

The phase separation of aqueous polymer solutions is a widely used method for producing self-assembled, membraneless droplet protocells. Non-ionic synthetic polymers forming an two-phase system (ATPS) have been shown to reliably form protocells that, when equipped with biological materials, are useful applications such as analyte detection. Previous characterization ATPS-templated protocell did not investigate the effects its components on stability. Here we report diagram PEG 35k-Ficoll...

RNA toehold switches are a widely used class of molecule to detect specific "trigger" sequences, but their design, intended function, and characterization date leave it unclear whether they can function properly with triggers shorter than 36 nucleotides. Here, we explore the feasibility using standard 23-nucleotide truncated triggers. We assess crosstalk different significant homology identify highly sensitive trigger region where just one mutation from consensus sequence reduce switch...

Abstract Simultaneous detection of multiple analytes from a single sample (multiplexing), particularly when at the point need, can guide complex decision-making without increasing required volume or cost per test. Despite recent advances, multiplexing still typically faces critical limitation measuring only one type molecule assay platform – for example, small molecules nucleic acids. In this work, we address bottleneck with customizable that integrates cell-free expression (CFE)...

Abstract RNA regulators are powerful components of the synthetic biology toolbox. Here, we expand repertoire gene networks built from these by constructing a transcriptional negative autoregulation (NAR) network out small RNAs (sRNAs). NAR motifs core natural genetic networks, and known for reducing response time steady state signal noise. Here use cell-free transcription-translation (TX-TL) reactions computational model to design prototype sRNA constructs. Using parameter sensitivity...

ABSTRACT The One-Pot PURE ( P rotein synthesis U sing R ecombinant E lements) system simplifies the preparation of traditional systems by co-culturing and purifying 36 essential proteins for gene expression in a single step, thereby improving accessibility affordability widespread laboratory adoption customization. However, replicating this protocol to match productivity can take considerable time effort due uncharacterized variability system’s biochemical composition. In work, we observed...

Large language models (LLMs) have demonstrated emergent capabilities across diverse reasoning tasks via popular Chains-of-Thought (COT) prompting. However, such a simple and fast COT approach often encounters limitations in dealing with complicated problems, while thorough method, which considers multiple pathways verifies each step carefully, results slower inference. This paper addresses the challenge of enabling LLMs to autonomously select between slow inference methods, thereby...

Ginsenoside compound K (CK) is one of the major metabolites bioactive ingredients in Panax ginseng, which presents excellent bioactivity and regulates expression important proteins. In this work, effects CK on G-quadruplexes (G4s) were quantitatively analyzed presence absence their complementary sequences. was demonstrated to facilitate formation G4s, increase quantity G4s competition with duplex. Thermodynamic experiments suggested that electrostatic interactions for G4 stabilization by CK....